title: Rapid lysis of gram-positive cocci for pulsed-field gel electrophoresis using achromopeptidase.
abstract: To adapt pulsed-field gel electrophoresis (PFGE) to a busy clinical laboratory, we evaluated the use of achromopeptidase (ACP) as the sole agent for lysis of a variety of gram-positive cocci. Growth in an appropriate broth media was adjusted to a 3.0 McFarland density before aliquotting for washing and plug preparation. ACP incorporated into the agarose plugs, or added to the buffer solution, lysed cells in < 1 h when suspended in a Tris-NaCl buffer. Two 30-min washes in a Tris-EDTA buffer were sufficient to remove cellular debris without additional deproteination. Final PFGE results were obtained in approximately 48 h, including the initial broth subculturing.
authors.affiliation: Associated Regional and University Pathologists, Inc., Salt Lake City, Utah, USA.
authors.forename: R B
authors.initials: RB
authors.lastname: Leonard
authors.forename: K C
authors.initials: KC
authors.lastname: Carroll
mesh_terms: D015373:Bacterial Typing Techniques; D001433:Bacteriolysis / Q000187:drug effects*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D016983:Enterococcus / Q000502:physiology; D005843:Geography; D006095:Gram-Positive Cocci / Q000502:physiology*; D012697:Serine Endopeptidases / Q000494:pharmacology*; D013211:Staphylococcus aureus / Q000502:physiology; D013296:Streptococcus pneumoniae / Q000502:physiology; D013297:Streptococcus pyogenes / Q000502:physiology
chemical_list: D012697:Serine Endopeptidases; C036813:lysyl endopeptidase